Isolation and characterization of a self‐sufficient one‐domain protein

Abstract

Earthworms have been shown to accumulate trace elements in general, and particularly high amounts of metal ions such as cadmium, copper and zinc. The earthworm's response to metal contamination has been linked to the induction and expression of metallothionein (MT) proteins, a detoxification strategy analogous to that found in other biological systems. The present study focuses on an inducible Cd‐MT isolated from the compost‐dwelling brandling worm Eisenia foetida (Savigny). A full characterization of the protein (including protein induction, MT cDNA, amino‐acid sequence and metal stoichiometry) revealed a new dimension of knowledge to the molecular genetic information available to date. Whereas the elucidated cDNA codes for a putative protein which possesses 80 amino‐acid residues, the characterized protein bears only 41 amino acids. The isolated product has evidently attained its size and shape by cleavage near the N‐terminal site and at the linker region between the two putative metal‐binding domains of the translated product, yielding a small MT moiety which contains 12 Cys residues (including one triple Cys‐motif) binding four cadmium ions. It can be shown that the isolated MT molecule represents a self‐sufficient one‐domain MT which is stable in vitro . The isolation of the single‐domain MT peptide raises the question about the method of formation and significance in vivo of such small MT moieties from tissues of E. foetida and possibly other terrestrial invertebrates. In this respect, two hypotheses are discussed: firstly, the possibility of formation of small MT peptides due to enzymatic cleavage of the intact protein during the process of preparation and isolation; and secondly, the possibility of deliberate post‐translational processing of the translated gene product to yield functional one‐domain MT moieties.